Fluorescence lifetime imaging of DAPI-stained nuclei as a novel diagnostic tool for the detection and classification of B-cell chronic lymphocytic leukemia

Gilad Yahav, Abraham Hirshberg, Ophira Salomon, Ninette Amariglio, Luba Trakhtenbrot, Dror Fixler

Research output: Contribution to journalArticlepeer-review

Abstract

B-cell chronic lymphocytic leukaemia (B-CLL) and B-cell precursor acute lymphoblastic leukaemia (B-ALL) are the most common type of leukaemia in adults and children, respectively. Today, fluorescence in situ hybridization (FISH) is the standard for detecting chromosomal aberrations that reflect adverse and favorable outcome. This study revealed a new, simple, and fast diagnostic tool to detect pathological cells by measuring and imaging the fluorescence lifetime (FLT) using FLT imaging microscopy (FLIM) of the peripheral blood (PB) cells of B-CLL samples that were labeled with the DNA binder, DAPI. The FLT of DAPI in healthy individuals was found to be 2.66 ± 0.12 ns. In contrast, PB cells of B-CLL and BM cells of B-ALL patients were characterized by a specific group distribution of the FLT values. The FLT of DAPI was divided into four subgroups, relative to 2.66 ns: short+, normal, prolonged, and prolonged+. These alterations could be related to different chromatin arrangements of B-CLL and B-ALL interphase nuclei. Notably, extremely long FLT of nuclear DAPI correlate with the presence of extra chromosome 12, while moderate increases compared to normal characterize the deletion of p53. Such correlations potentially enable a FLT-based rapid automatic diagnosis and classification of B-CLL even when the frequency of genetic and chromosomal abnormalities is low.

Original languageEnglish
Pages (from-to)644-652
Number of pages9
JournalCytometry. Part A : the journal of the International Society for Analytical Cytology
Volume89
Issue number7
DOIs
StatePublished - 1 Jul 2016

Keywords

  • B-cell chronic lymphocytic leukaemia
  • B-cell precursor acute lymphoblastic leukaemia
  • bone marrow
  • fluorescence in situ hybridization
  • fluorescence lifetime
  • fluorescence lifetime imaging microscopy

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